This study was designed to investigate inhibitory effects and possible mechanisms of snake venom tripeptide (pENW) on platelet adhesion in order to promote the development of a novel anti-platelet therapy. To study the inhibitory effects of pENW on platelet adhesion, washed platelets pre-incubated with pENW (116.5-466.2 μmol x L(-1)) were used to test the ability of platelet adhesion to fibrinogen. Effect of pENW on fibrin clot retraction was also tested. Effect of pENW on platelets viability was tested by MTT assay. Effect of pENW on reactive-oxygen species (ROS) levels of platelet was studied by flow cytometry assay. Calcium mobilization in Fura-2/AM-loaded platelets was monitored with a spectrofluorimeter. Cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP), thromboxane A2 (determined as its metabolite thromboxane B2) were measured using enzyme immunoassay kits. Akt, ERK and p38 phosphorylation were tested by Western blot. The results showed that pENW inhibited platelet adhesion and fibrin clot retraction in a concentration-dependent manner without cytotoxicity. Intracellular cGMP and cAMP in both resting and thrombin-activated platelets were increased by pENW. In addition, pENW attenuated intracellular Ca2+ mobilization and TXA2 production in platelets stimulated by thrombin. As shown by Western blot assay, Akt, ERK and p38 phosphorylation in thrombin-induced platelet were attenuated by pENW. However, inhibitory effects of pENW had nothing to do with ROS. Thus, pENW exhibited a significant inhibition on platelet adhesion to fibrinogen, which means pENW could block the first step of thrombosis as while as retard the more stable clot formation. The mechanisms of pENW on inhibition platelet adhesion might be related to instant regulations, such as protein kinases.
Blood Platelets ; drug effects ; Blotting, Western ; Calcium ; metabolism ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Flow Cytometry ; Phosphorylation ; Platelet Aggregation ; drug effects ; Reactive Oxygen Species ; metabolism ; Snake Venoms ; chemistry ; Thromboxane A2 ; metabolism ; Thromboxane B2 ; metabolism

OBJECTIVETo investigate the clinicopathologic feature, immunophenotype and differential diagnosis of cutaneous Rosai-Dorfman disease (CRDD).

METHODSClinical manifestation, morphologic features and immunohistochemical staining were studied in 8 cases of CRDD.

RESULTSAll 8 patients presented with multiple papules, nodules and/or coalescent patches or plaques distributing over the extremities or trunk, without lymphadenopathy or other systemic abnormalities. Microscopically, the lesions were located intradermally and/or subcutaneously. CRDD was characterized by the presence of S-100 positive histiocytic cells exhibiting emperipolesis, accompanying with infiltration of mixed inflammatory cells. Fibrosis, somewhere in vague storiform pattern due to stromal responses, with distribution of individual neutrophil microabscess was seen in cases with a long course of illness. Dilated vascular spaces in dermis containing numerous large typical histiocytes were seen in 2 cases.

CONCLUSIONSCRDD is a benign, persistent proliferative disease of histiocytes. Systemic involvement is rare, outcome favorable. It should be differentiated from other types of histiocytosis, dermatofibrosarcoma protuberans, xanthoma and lymphoproliferative disorders. Immunohistochemical staining for S-100 protein and CD68 is helpful in making a correct diagnosis.

Aged ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Diagnosis, Differential ; Female ; Histiocytosis, Sinus ; metabolism ; pathology ; surgery ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Prognosis ; S100 Proteins ; metabolism ; Skin Diseases ; metabolism ; pathology ; surgery

OBJECTIVETo study the clinicopathologic features, immunophenotype and prognosis of primary cutaneous anaplastic large cell lymphoma (CALCL).

METHODSHistopathologic evaluation and immunohistochemical study by Envision method were carried out in 44 archival cases of CALCL. The clinical information and follow-up data were analyzed.

RESULTSThe patients presented with skin nodules, masses or plaques, sometimes associated with ulceration. The commonest sites of involvement were the extremities. Follow-up data were available in 39 patients. The overall survival rate was 87.2% (34/39). Disease relapses were detected in 46.2% (18/39) of the patients. Statistical analysis indicated that patients older than 50 years of age or with no less than two involved anatomic sites were more likely to have disease relapses (P < 0.05). Histologically, 31 cases were classified as common variant, 6 cases as small cell variant and 7 cases as neutrophil/eosinophil-rich variant. Immunohistochemical study showed that the rates of expression of CD30, CD45, CD45RO, CD43, CD3, cytotoxic protein and epithelial membrane antigen were 100% (44/44), 91.2% (31/34), 82.6% (19/23), 94.7% (18/19), 70.0% (28/40), 73.3% (22/30) and 31.8% (7/22), respectively. The CD4(+)/CD8(-), CD4(-)/CD8(+) and CD4(-)/CD8(-) immunophenotypes were found in 58.3% (21/36), 22.2% (8/36) and 19.4% (7/36) of the CALCL cases, respectively. Only one case (3.7%) expressed CD56.

CONCLUSIONSCALCL is a form of low-grade primary cutaneous T-cell lymphoma with a wide spectrum of clinicopathologic pattern. Special variants of CALCL should not be confused with other types of cutaneous lymphomas and inflammatory lesions. CALCL patients older than 50 years of age or with no less than two involved anatomic sites are more likely to have disease relapses.

Adult ; Age Factors ; Aged ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Immunophenotyping ; Ki-1 Antigen ; metabolism ; Lymphoma, Large-Cell, Anaplastic ; metabolism ; pathology ; Lymphoma, Primary Cutaneous Anaplastic Large Cell ; drug therapy ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Proportional Hazards Models ; Skin Neoplasms ; drug therapy ; metabolism ; pathology ; Survival Rate ; Young Adult

AIMTo evaluate the best individualized prostate biopsy strategies for Chinese patients with suspected prostate cancer.

METHODSThe present study included 221 Chinese patients who underwent transrectal ultrasound guided prostate biopsies for the first time. All patients underwent the same 10-core biopsy protocol. In addition to the Hodge sextant technique, four more biopsies were obtained from the base and middle regions of bilateral peripheral zones. The differences between 10-core and sextant strategies in cancer detection among patients with different prostate specific antigen (PSA) levels were evaluated. The relationship between PSA level, number of positive biopsy cores and organ-confined cancer rate in prostate cancer patients was also analyzed.

RESULTSThe overall prostate cancer detection rate was 40.7% in the 221 patients. The 10-core strategy increased cancer detection by 6.67% (6/90) in our patients (P < 0.05). The increased cancer detection rates decreased significantly when the patient PSA level increased from 0-20 ng/mL to 20.1-50 ng/mL and > 50 ng/mL (P < 0.01). The number of positive biopsy cores in prostate cancer patients increased significantly with increasing patient PSA level (P < 0.01). The rate of organ-confined prostate cancer decreased significantly with increasing patient PSA level (P < 0.01).

CONCLUSIONThe extended 10-core strategy is recommended for Chinese patients with PSA = or < 20 ng/mL and the sextant strategy is recommended for those with PSA > 50 ng/mL. For patients with PSA ranging from 20.1 ng/mL to 50 ng/mL, the 10-core strategy should be applied in patients with life expectancy = or > 10 years and the sextant strategy should be applied in those with life expectancy < 10 years.

Aged ; Aged, 80 and over ; Biopsy, Needle ; China ; Humans ; Male ; Middle Aged ; Prostate ; pathology ; Prostate-Specific Antigen ; blood ; Prostatic Neoplasms ; blood ; pathology

PURPOSE: Cefodizime is a new third-generation cephalosporin which has a structure and immunomodutation properties similar to cefotaxime. Various studies on cefodizime have demonstrated the direct eradication of bacteria in cooperation with the host defense mechanism, particularly with phagocytosis. We evaluated the effects of cefodizime on the phagocytosis of COS-1 cells transfected with FcgammaRI/gammagamma or FcgammaRIIA cDNA. METHODS: Phagocytosis was measured using the in vitro COS-1 cell modeling system according to Schreiber's method. COS-1 cells, which lack endogenoous Fcgammareceptors but have phagocytic potential, were transfected with either FcgammaRI/gammagammaor FcgammaRIIA cDNA. COS-1 cells, as target cells, were treated with antibiotics for 1 or 24 hours and incubated for 30 min with IgG coated sheep RBCs. Adhered IgG coated sheep RBCs were removed after brief exposure to hypotonic phosphate buffered saline. Phagocytosis index (PI) was calculated as the number of ingested RBCs per 100 phagocytic cells after wright-Giemsa staining. RESULTS: COS-1 cells tranfected with FcgammaR (either FcgammaRI/gammagamma or FcgammaRIIA cDNA) showed the phagocytic activity against IgG coated sheep RBC, while untransfected COS-1 cells did not. After treatment with cefodizime, phagocytic activity of FcgammaRI/gammagammacDNA transfected COS-1 cells was significantly increased, while that of FcgammaRIIA cDNA transfected COS-1 cells did not. Marked enhancement of phagocytosis of COS-1 cells was observed after treatment with cefodizime, but was not observed with ceftriaxone or moxalactam. CONCLUSION: Cefodizime showed marked enhancement of phagocytic activity of FcgammaR transfected COS-1 cells. FcgammaRI seems to play an important role in the enhancement of phagocytosis. Further studies will be required.
Animals ; Anti-Bacterial Agents ; Bacteria ; Cefotaxime ; Ceftriaxone ; COS Cells ; DNA, Complementary ; Immunoglobulin G ; Moxalactam ; Phagocytes ; Phagocytosis* ; Sheep

PURPOSE: The prevalence of atopic diseases has been increasing remarkably. The less frequent opportunities for infection early in life, especially mycobacteria exposure, parallel this higher prevalence of atopic diseases. Bacille Calmette-Gu rin (BCG), a potent inducer of Th1 immune response, has been suggested to suppress Th2 response which is known to mediate IgE-mediated atopic disorders. This study was done to investigate whether there is any relation between the number of BCG scars and the prevalence of atopic disorders in early childhood. METHODS: We surveyed 393 parents with a children who were given percutaneous multi- puncture BCG vaccination within four weeks after birth. The main questions concerned the past history and present illness of physician-diagnosed atopic diseases (atopic dermatitis, bronchial asthma, and allergic rhinitis), the number of BCG scars (range; 0-18), and potential confounders such as gender, parental atopy, maternal smoking and environmental cofactors. The prevalence rate of each atopic disease was measured and analysed according to the number of BCG scars. RESULTS: Each prevalence rate was 18.1% for atopic dermatitis, 9.4% for bronchial asthma, 14.6% for allergic rhinitis, and 32.3% for any of them. All of them had received BCG vaccination during the first four weeks of life. The children with 15 or more BCG scars had a significantly lower prevalence of any atopic disease (22/99, 22.2%) as compared to those with four scars or less (51/125, 40.8%) by simple regression analysis. (P value=0.02) But this association was not significant after controlling for potential confounders. (P value= 0.26) CONCLUSION: This survey demonstrated a weak relation between a larger number of BCG scars and less atopy development at early childhood. But the relation was not so significant. Further studies are needed.
Asthma ; Child ; Cicatrix* ; Dermatitis ; Dermatitis, Atopic ; Humans ; Mycobacterium bovis* ; Parents ; Parturition ; Prevalence ; Punctures ; Rhinitis ; Smoke ; Smoking ; Vaccination

OBJECTIVETo raise the vigilance not to believe easily the diagnosis of a primary branchial cleft carcinoma.

METHODSFour cases of cystic metastatic squamous cell carcinoma in the neck misdiagnosed as branchiogenic carcinoma from 1993 to 2002 in our hospital were analyzed retrospectively.

RESULTSThe primary sites of these 4 cases were later discovered, 2 in the aryepiglottic fold, 1 in faucial tonsil and 1 in the skin of the head, respectively. The discovery of the primary sites ranged from the day of initial surgery to 41 months.

CONCLUSIONNone of the cases reviewed in this study was a branchiogenic carcinoma. Therefore, the diagnosis of a primary branchial cleft carcinoma requires the fulfillment of strict criteria both clinically and pathologically.

Adult ; Branchioma ; diagnosis ; Carcinoma, Squamous Cell ; diagnosis ; secondary ; Diagnostic Errors ; Epiglottis ; Female ; Head and Neck Neoplasms ; diagnosis ; secondary ; Humans ; Laryngeal Neoplasms ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Tonsillar Neoplasms ; pathology

BACKGROUNDEndothelial dysfunction is thought to be critical events in the pathogenesis of Alzheimer's disease (AD). Endothelial progenitor cells (EPCs) have provided insight into maintaining and repairing endothelial function. To study the relation between EPCs and AD, we explored the number of circulating EPCs in patients with AD.

METHODSA total of 104 patients were recruited from both the outpatients and inpatients of the geriatric neurology department at General Hospital, Tianjin Medical University. Consecutive patients with newly diagnosed AD (n = 30), patients with vascular dementia (VaD, n = 34), and healthy elderly control subjects with normal cognition (n = 40) were enrolled after matching for age, gender, body mass index, medical history, current medication and Mini Mental State Examination. Middle cerebral artery flow velocity was examined with transcranial Doppler. Endothelial function was evaluated according to the level of EPCs, and peripheral blood EPCs was counted by flow cytometry.

RESULTSThere were no significant statistical differences of clinical data in AD, VaD and control groups (P > 0.05). The patients with AD showed decreased CD34-positive (CD34(+)) or CD133-positive (CD133(+)) levels compared to the control subjects, but there were no significant statistical differences in patients with AD. The patients with AD had significantly lower CD34(+)CD133(+) EPCs (CD34 and CD133 double positive endothelial progenitor cells) than the control subjects (P < 0.05). In the patients with AD, a lower CD34(+)CD133(+) EPCs count was independently associated with a lower Mini-Mental State Examination score (r = 0.514,P = 0.004). Patients with VaD also showed a significant decrease in CD34(+)CD133(+) EPCs levels, but this was not evidently associated with the Mini-Mental State Examination score. The changes of middle cerebral artery flow velocity were similar between AD and VaD. Middle cerebral artery flow velocity was decreased in the AD and VaD groups and significantly lower than the normal control group (P < 0.01). There was no significant difference of the blood flow velocity between the AD and VaD patients (P > 0.05).

CONCLUSIONSThe results provided evidence that patients with AD have reduced circulating EPCs. Endothelial function is impaired in patients with AD and vascular factors have a role in the pathogenesis of AD. CD34(+)CD133(+) EPCs may be a novel biomarker of AD dementia.

AC133 Antigen ; Aged ; Alzheimer Disease ; metabolism ; pathology ; Antigens, CD ; metabolism ; Antigens, CD34 ; metabolism ; Dementia, Vascular ; metabolism ; pathology ; Endothelial Cells ; cytology ; metabolism ; Female ; Glycoproteins ; metabolism ; Humans ; Male ; Peptides ; metabolism ; Stem Cells ; cytology ; metabolism

OBJECTIVETo investigate diagnostic histopathology and ultrastructure features of primary pulmonary cryptococcosis (PC).

METHODSClinical data and pathologic findings of 27 cases of PC were retrospectively reviewed, light and electron microscopic evaluations and histochemistry stain studies were performed.

RESULTSThe specimens consisted of 2 fine-needle aspiration lung biopsies and 25 cases of open lung biopsies. Cryptococcosis granuloma formation was identifiable by histopathological examination in 25 of 27 cases, with gum-like lesion and fungi in the remaining 2 cases. The detection rates of cryptococcus neoformans (CN) by mucicarmine (MC), periodic acid-Schiff (PAS), alcian blue (AB) and Grocott methenamine-silver (GMS) were 87.0% (20/23), 100% (27/27), 66.7% (18/27), and 100% (23/23) respectively. Under the electron microscope, most CN had a simple structure with a few organelles. The capsule was seen in all organisms. A percentage of the organisms showed nuclei, nucleoli, mitochondria and vacuoles. The detection rate of CN by EM was 91.7% (11/12).

CONCLUSIONSThe clinical manifestation and imaging of PC are nonspecific for PC. Lung biopsy is the major diagnostic modality. The detection rate by electron microscopy was quite high. Therefore, a correct diagnosis of pulmonary cyrptococcosis should rely on the combination of histopathological evaluation, histochemistry staining and/or electron microscopic examination.

Adult ; Aged ; Biopsy, Fine-Needle ; Cryptococcosis ; microbiology ; pathology ; Cryptococcus neoformans ; isolation & purification ; ultrastructure ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Lung ; pathology ; ultrastructure ; Lung Diseases, Fungal ; classification ; microbiology ; pathology ; Male ; Microscopy, Electron ; Middle Aged ; Retrospective Studies

OBJECTIVETo observe the effect of intracerebroventricular injection of rAAV-HIF-1α on hippocampal neuronal apoptosis in a rat model of Alzheimer disease (AD).

METHODSThirty-two male SD rats (250-300 g) were randomized into 4 groups (n=8), including the normal control group without any treatment, AD model group with right intracerebroventricular injection of 2 µl Aβ25-35 (10 mg/m1), sham-operated group with right intracerebroventricular injection of 2 µl normal saline, and AD+ rAAV-HIF-1α group with right intracerebroventricular injection of 10 µl rAAV-HIF-1a (1×10¹² v.g./m1) one week after Aβ25-35 injection. The rats were sacrificed to detect the expression of HIF-1α and apoptosis of hippocampal neurons 5 weeks after Aβ25-35 or saline injection.

RESULTSWestern blotting showed that the expression of HIF-1α was significantly higher in AD+rAAV-HIF-1α group (451.59±34.39) than in normal control group (229.05±41.28) and sham-operated group (216.29±37.08) (P<0.05) without significant difference between the latter two groups. The apoptotic ratio of the hippocampal neurons was significantly higher in AD model group ([19.49±2.59]%) than in normal control group ([5.41±0.75]%) and sham-operated group ([5.28±0.66]%) in (P<0.05), and intracerebroventricular injection of rAAV-HIF-1α resulted in a significant reduction of the apoptotic ratio in the AD rats ([12.07±2.06]%) (P<0.05).

CONCLUSIONIntracerebroventricular injection of rAAV-HIF-1α can inhibit hippocampal neuronal apoptosis in the rat model of AD.

Alzheimer Disease ; metabolism ; therapy ; Animals ; Apoptosis ; Disease Models, Animal ; Hippocampus ; cytology ; Hypoxia-Inducible Factor 1, alpha Subunit ; administration & dosage ; genetics ; metabolism ; Lateral Ventricles ; Male ; Neurons ; drug effects ; pathology ; Rats ; Rats, Sprague-Dawley